The long-term goal of this project is to develop novel, broad-spectrum antibiotics against both biowarfare agents and common drug resistant pathogens. The potential use of engineered, multi-drug-resistant strains as agents of bioterrorism, and the current spread of conventional drug resistant pathogens necessitate the development of novel antibiotics. Most antibiotics in use today have been obtained from less than 1% of microbial species due to the inability to culture the vast majority of species by current methods. The remaining 99% of species represent essentially an unlimited diversity of microorganisms to discover novel antibiotics. In this Phase I project we aim to develop an industrial-scale methodology to grow and screen previously uncultivated bacteria, including Actinobacteria, and obtain proof-of-principle for production of novel broad-spectrum antibiotics from this untapped source of secondary metabolites. To access the "uncultivable" species, we will capitalize on our proprietary method based on in situ cultivation of environmental microorganisms, followed by their domestication in vitro. To achieve this goal, we will develop the method into a unique high throughput technology platform for massive cultivation and identification of new antibiotics from "uncultivable" microorganisms. The specific aims of the project are: Aim 1: Develop ichip, the cultivation device combining hundreds of miniature diffusion chambers as a technology for a rapid, high throughput, in situ cultivation and domestication of "uncultivable" microorganisms;Aim 2: Identify species with broad spectrum antimicrobial activities using a panel of multidrug resistant pathogens and biowarfare agents;Aim 3: Investigate the chemical novelty of the discovered activities. The end product will be a pipeline for discovery of novel antimicrobials, which we will fully utilize in Phase II to develop novel broad spectrum compounds. The annual market for antimicrobials is $25 billion, and several broad-spectrum drugs have sales in excess of $1 billion. The large-scale screening in Phase II will produce a variety of candidate molecules, which will undergo detailed validation, ultimately resulting in new antibiotic products targeting this market. PUBLIC HEALTH RELEVANCE: The rapid acquisition of resistance of bacterial pathogens in both community and hospital based infections coupled with the decreased investment in novel antibiotic development by large pharmaceutical companies represents an impending public health crisis. The purpose of this project is to develop a new technology to discover novel antibiotics. These new antibiotics will provide the public with a stronger defense whether the threat is from conventional or bioterror bacterial pathogens.